The aim of the proposed project is to elucidate the molecular mechanisms of gene action in man through biochemical approach. The project consists of the following studies: 1) Determination of complete amino acid sequence of normal human glucose 6-phospate dehydrogenase and phosphoglyceratekinase, and elucidation of amino acid substitutions of variant enzymes. Because of the existince of many abnormal enzymes associated with hemolytic disorders, these studies, together with the three dimensional structure of the enzymes, molecular pathology of these enzyme abnormalities will be advanced. Screening and characterization of new variant enzymes will be carried out. 2) Determination of complete amino acid and carbohydrate sequence of normal alpha 1-protease inhibitor. Amino and substitution and carbohydrate abnormality of variant inhibitors will be undertaken. The knowledge obtaind by these studies may lead to an understanding of protease inhibitor abnormalities associated with emphysema. 3) Purification and characterization of glycosyltransferases involved in synthsis of blood group substances. Studies of structure and immunologic homology of N-acetylgalactosaminyltransferase, galactosyltransferase and other trasferases in human blood will lead to an understanding of underlying genetic mechanisms of expression of blood types. 4) Purification and characterization of mannosidase, abnormal pseudocholinesterase and atypical liver alcohol dehydrogenase related to genetic disorders.